Genetic Engineering: A Revolution in Molecular Biology - Outline

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11 Δεκ 2012 (πριν από 4 χρόνια και 8 μήνες)

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1
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1

Chapter
10

Genetic Engineering: A Revolution in Molecular Biology


Chapter Outline


10.1.

Basic Elements and Applications of Genetic Engineering


A. Genetics is considered
basic
science


B. Genetic engineering is an
applied

science

10.2.

Tools and Techniques of
Genetic Engineering

A. DNA: The raw material


1. Enzymes for dicing, splicing, and reversing nucleic acids



a. Restriction endonucleases




i. Palindromes







ii. EcoRI from
E. coli

and HindIII from
Haemophilus influenzae

iii. Restriction fra
gments and restriction fragment length








polymorphisms (RFLPS)



b. Ligase



c. Reverse transcriptase



d. Complementary DNA (cDNA)


2. Analysis of DNA



a. Gel electrophoresis


3. Nucleic acid hybridization and probes



a. Gene probes



b. Southern blot method



c. FISH





4. Methods used to size, synthesize, and sequence DNA

a.

DNA sequencing: Determining the exact genetic code







i. Sanger method


5. Polymerase chain reaction: A molecular Xerox machine for DNA



a. Primers



b. DN
A polymerases







i.

Thermus aquaticus and Thermococcus litoralis






c. Steps in PCR cycle:









i. Denaturation







ii. Priming







iii. Extension

10.3.

Methods in Recombination DNA Technology: How to Imitate Nature
-
Or to “One
-
Up” It



A. In
troduction




1. Recombinant DNA technology





a. Clones





b. Vectors





c. Cloning hosts



B
. Technical
a
spects of
r
ecombinant DNA and
g
ene
cloning


1. Characteristics of cloning vectors



a. Charon phage



b. Cosmids



c. Artificial chromosomes


2
. Characteristics of cloning hosts



a.
E. coli



b.
Saccharomyces cerevesiae



1
-
2



c
.

Bacillus subtilis




C
. Construction of a
recombinant
,
i
nsertion into a
c
loning
h
ost, and
g
enetic
e
xpression


10.4.

Biochemical Products of Recombinant DNA Technology



A.

Human growth hormone



B. Insulin




C. Clotting factor VIII

10.5.

Genetically Modified Organisms

A. Recombinant microbes: Modified bacteria and viruses


1.
Pseudomonas syringae

and Frostban





2.
Pseudomonas fluorescens, Bacillus thuringiensis

and

insecticide genes

B. Transgenic plants: Improving crops and foods


1.
Agrobacterium




C. Transgenic animals: Engineering embryos





1. Pharming

10.6.

Genetic Treatments: Introducing DNA into the Body

A. Gene therapy



1. Adenosine deaminase (ADA)
deficiency


2. SCID

B. DNA technology as genetic medicine


1. Antisense DNA: Targeting messenger RNA


10.7.

Genome Analysis: Maps, Fingerprints, and Family Trees


A. Genome mapping and screening: An atlas of the genome



1. Locus and alleles



2. Li
nkage, physical and sequence maps





3. Genomics and bioinformatics


B. DNA fingerprinting: A unique picture of a genome





1. Single nucleotide polymorphism





2. Markers






a. Variable number of tandem repeats (VNTRs)






b. Microsatellite po
lymorphisms





3. Microarray analysis